title: Cell nucleus targeting for living cell extraction of nucleic acid associated proteins with intracellular nanoprobes of magnetic carbon nanotubes.
abstract: Since nanoparticles could be ingested by cells naturally and target at a specific cellular location as designed, the extraction of intracellular proteins from living cells for large-scale analysis by nanoprobes seems to be ideally possible. Nucleic acid associated proteins (NAaP) take the crucial position during biological processes in maintaining and regulating gene structure and gene related behaviors, yet there are still challenges during the global investigation of intracellular NAaP, especially from living cells. In this work, a strategy to extract intracellular proteins from living cells with the magnetic carbon nanotube (oMWCNT@Fe3O4) as an intracellular probe is developed, to achieve the high throughput analysis of NAaP from living human hepatoma BEL-7402 cells with a mass spectrometry-based proteomic approach. Due to the specific intracellular localization of the magnetic carbon nanotubes around nuclei and its strong interaction with nucleic acids, the highly efficient extraction was realized for cellular NAaP from living cells, with the capability of identifying 2383 intracellular NAaP from only ca. 10,000 living cells. This method exhibited potential applications in dynamic and in situ analysis of intracellular proteins.
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zhengyan
authors.initials: Z
authors.lastname: Hu
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Qin
authors.forename: Fangjie
authors.initials: F
authors.lastname: Liu
authors.forename: Kai
authors.initials: K
authors.lastname: Cheng
authors.forename: Ren'an
authors.initials: R
authors.lastname: Wu
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D002470:Cell Survival; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D005290:Ferric Compounds / Q000737:chemistry; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D059346:Magnets; D015335:Molecular Probes / Q000737:chemistry*; D037742:Nanotubes, Carbon / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D004268:DNA-Binding Proteins; D005290:Ferric Compounds; D015335:Molecular Probes; D037742:Nanotubes, Carbon; D016601:RNA-Binding Proteins; C000499:ferric oxide